Comprehensive Analysis of 13C6 Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts

The bioenergetics of the vast majority of terrestrial mammals evolved to consuming glucose (Glc) for energy production under regular atmosphere (about 21% oxygen). However, some vertebrate species, such as aquatic turtles, seals, naked mole rat, and blind mole rat, Spalax, have ad...

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Autores principales:	Dmitry Miskevich, Anastasia Chaban, Maria Dronina, Ifat Abramovich, Eyal Gottlieb, Imad Shams
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	metabolome hypoxia proline cycle GSH adaptation bioenergetics Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/0162c05384494195a76d71f67c526c6a
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spelling	oai:doaj.org-article:0162c05384494195a76d71f67c526c6a2021-11-25T18:20:32ZComprehensive Analysis of <sup>13</sup>C<sub>6</sub> Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts10.3390/metabo111107342218-1989https://doaj.org/article/0162c05384494195a76d71f67c526c6a2021-10-01T00:00:00Zhttps://www.mdpi.com/2218-1989/11/11/734https://doaj.org/toc/2218-1989The bioenergetics of the vast majority of terrestrial mammals evolved to consuming glucose (Glc) for energy production under regular atmosphere (about 21% oxygen). However, some vertebrate species, such as aquatic turtles, seals, naked mole rat, and blind mole rat, <i>Spalax</i>, have adjusted their homeostasis to continuous function under severe hypoxic environment. The exploration of hypoxia-tolerant species metabolic strategies provides a better understanding of the adaptation to hypoxia. In this study, we compared Glc homeostasis in primary <i>Spalax</i> and rat skin cells under normoxic and hypoxic conditions. We used the targeted-metabolomics approach, utilizing liquid chromatography and mass spectrometry (LC-MS) to track the fate of heavy Glc carbons (<sup>13</sup>C<sub>6</sub> Glc), as well as other methodologies to assist the interpretation of the metabolic landscape, such as bioenergetics profiling, Western blotting, and gene expression analysis. The metabolic profile was recorded under steady-state (after 24 h) of the experiment. Glc-originated carbons were unequally distributed between the cytosolic and mitochondrial domains in <i>Spalax</i> cells compared to the rat. The cytosolic domain is dominant apparently due to the hypoxia-inducible factor-1 alpha (HIF-1α) mastering, since its level is higher under normoxia and hypoxia in <i>Spalax</i> cells. Consumed Glc in <i>Spalax</i> cells is utilized for the pentose phosphate pathway maintaining the NADPH pool, and is finally harbored as glutathione (GSH) and UDP-GlcNAc. The cytosolic domain in <i>Spalax</i> cells works in the semi-uncoupled mode that limits the consumed Glc-derived carbons flux to the tricarboxylic acid (TCA) cycle and reduces pyruvate delivery; however, it maintains the NAD<sup>+</sup> pool via lactate dehydrogenase upregulation. Both normoxic and hypoxic mitochondrial homeostasis of Glc-originated carbons in <i>Spalax</i> are characterized by their massive cataplerotic flux along with the axis αKG→Glu→Pro→hydroxyproline (HPro). The product of collagen degradation, HPro, as well as free Pro are apparently involved in the bioenergetics of <i>Spalax</i> under both normoxia and hypoxia. The upregulation of 2-hydroxyglutarate production detected in <i>Spalax</i> cells may be involved in modulating the levels of HIF-1α. Collectively, these data suggest that <i>Spalax</i> cells utilize similar metabolic frame for both normoxia and hypoxia, where glucose metabolism is switched from oxidative pathways (conversion of pyruvate to Acetyl-CoA and further TCA cycle processes) to (i) pentose phosphate pathway, (ii) lactate production, and (iii) cataplerotic pathways leading to hexosamine, GSH, and HPro production.Dmitry MiskevichAnastasia ChabanMaria DroninaIfat AbramovichEyal GottliebImad ShamsMDPI AGarticlemetabolomehypoxiaproline cycleGSHadaptationbioenergeticsMicrobiologyQR1-502ENMetabolites, Vol 11, Iss 734, p 734 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	metabolome hypoxia proline cycle GSH adaptation bioenergetics Microbiology QR1-502
spellingShingle	metabolome hypoxia proline cycle GSH adaptation bioenergetics Microbiology QR1-502 Dmitry Miskevich Anastasia Chaban Maria Dronina Ifat Abramovich Eyal Gottlieb Imad Shams Comprehensive Analysis of <sup>13</sup>C<sub>6</sub> Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts
description	The bioenergetics of the vast majority of terrestrial mammals evolved to consuming glucose (Glc) for energy production under regular atmosphere (about 21% oxygen). However, some vertebrate species, such as aquatic turtles, seals, naked mole rat, and blind mole rat, <i>Spalax</i>, have adjusted their homeostasis to continuous function under severe hypoxic environment. The exploration of hypoxia-tolerant species metabolic strategies provides a better understanding of the adaptation to hypoxia. In this study, we compared Glc homeostasis in primary <i>Spalax</i> and rat skin cells under normoxic and hypoxic conditions. We used the targeted-metabolomics approach, utilizing liquid chromatography and mass spectrometry (LC-MS) to track the fate of heavy Glc carbons (<sup>13</sup>C<sub>6</sub> Glc), as well as other methodologies to assist the interpretation of the metabolic landscape, such as bioenergetics profiling, Western blotting, and gene expression analysis. The metabolic profile was recorded under steady-state (after 24 h) of the experiment. Glc-originated carbons were unequally distributed between the cytosolic and mitochondrial domains in <i>Spalax</i> cells compared to the rat. The cytosolic domain is dominant apparently due to the hypoxia-inducible factor-1 alpha (HIF-1α) mastering, since its level is higher under normoxia and hypoxia in <i>Spalax</i> cells. Consumed Glc in <i>Spalax</i> cells is utilized for the pentose phosphate pathway maintaining the NADPH pool, and is finally harbored as glutathione (GSH) and UDP-GlcNAc. The cytosolic domain in <i>Spalax</i> cells works in the semi-uncoupled mode that limits the consumed Glc-derived carbons flux to the tricarboxylic acid (TCA) cycle and reduces pyruvate delivery; however, it maintains the NAD<sup>+</sup> pool via lactate dehydrogenase upregulation. Both normoxic and hypoxic mitochondrial homeostasis of Glc-originated carbons in <i>Spalax</i> are characterized by their massive cataplerotic flux along with the axis αKG→Glu→Pro→hydroxyproline (HPro). The product of collagen degradation, HPro, as well as free Pro are apparently involved in the bioenergetics of <i>Spalax</i> under both normoxia and hypoxia. The upregulation of 2-hydroxyglutarate production detected in <i>Spalax</i> cells may be involved in modulating the levels of HIF-1α. Collectively, these data suggest that <i>Spalax</i> cells utilize similar metabolic frame for both normoxia and hypoxia, where glucose metabolism is switched from oxidative pathways (conversion of pyruvate to Acetyl-CoA and further TCA cycle processes) to (i) pentose phosphate pathway, (ii) lactate production, and (iii) cataplerotic pathways leading to hexosamine, GSH, and HPro production.
format	article
author	Dmitry Miskevich Anastasia Chaban Maria Dronina Ifat Abramovich Eyal Gottlieb Imad Shams
author_facet	Dmitry Miskevich Anastasia Chaban Maria Dronina Ifat Abramovich Eyal Gottlieb Imad Shams
author_sort	Dmitry Miskevich
title	Comprehensive Analysis of <sup>13</sup>C<sub>6</sub> Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts
title_short	Comprehensive Analysis of <sup>13</sup>C<sub>6</sub> Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts
title_full	Comprehensive Analysis of <sup>13</sup>C<sub>6</sub> Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts
title_fullStr	Comprehensive Analysis of <sup>13</sup>C<sub>6</sub> Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts
title_full_unstemmed	Comprehensive Analysis of <sup>13</sup>C<sub>6</sub> Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts
title_sort	comprehensive analysis of <sup>13</sup>c<sub>6</sub> glucose fate in the hypoxia-tolerant blind mole rat skin fibroblasts
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/0162c05384494195a76d71f67c526c6a
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Comprehensive Analysis of <sup>13</sup>C<sub>6</sub> Glucose Fate in the Hypoxia-Tolerant Blind Mole Rat Skin Fibroblasts

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